NDLI: The Behaviour of Reduced, Alkylated and Native Proteins in a pH-Gradient LC System

Content Provider	Springer Nature Link
Author	Tran, Bao Pepaj, Miliam Lundanes, Elsa Greibrokk, Tyge
Copyright Year	2007
Abstract	Two dimensional (2D) liquid chromatography (LC) separations of proteins can be obtained faster and more automated than traditional 2D gel electrophoresis. Previously we have described a 2D LC method for separation of native proteins with separation according to pI by pH-gradient strong anion exchange (SAX) chromatography in the first dimension, and according to hydrophobicity by reversed phase chromatography in the second dimension. Since there are few literature reports on the combination of reduced/alkylated proteins and modern LC, a basic study of the chromatographic properties of a few reduced /alkylated proteins was undertaken with a pH-gradient SAX chromatographic system. Proteins where the disulfide groups were reduced, but not alkylated, were also included. The conditions that separated native proteins according to pI could not be used for neither reduced nor reduced/alkylated proteins. High concentrations of urea (4–8 M) were needed in the mobile phase in order to obtain good peak shapes. Addition of urea had an undesired impact on both the retention of the proteins and the pH gradient profile, with the effect that little correlation between reported pI values and elution pH was found. The conclusion was that proteins should be separated in the native state if good pI–pH correlations are important, and in the alkylated state with urea if other considerations are more important.
Starting Page	709
Ending Page	715
Page Count	7
File Format	PDF
ISSN	00095893
Journal	Chromatographia
Volume Number	66
Issue Number	9-10
e-ISSN	16121112
Language	English
Publisher	Vieweg Verlag
Publisher Date	2007-10-09
Publisher Place	Wiesbaden
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Ion-exchange chromatography pH-Gradient Urea Reduction and alkylation Plasma Measurement Science, Instrumentation Plant Sciences Biochemistry Pharmacy Organic Chemistry Analytical Chemistry
Content Type	Text
Resource Type	Article
Subject	Organic Chemistry Analytical Chemistry Biochemistry Clinical Biochemistry

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Refolding human lysozyme produced as an inclusion body by urea concentration and pH gradient ion exchange chromatography

Investigation of the ion-exchange behaviour of titania: Application as a packing material for ion chromatography

Retention behaviour of an homologous series of oligodeoxythymidilic acids using reversed-phase ion-pair chromatography

LC–MS–MS Determination of Olmesartan in Human Plasma

Gas chromatographic behaviour of urea herbicides

Simple and Sensitive Determination of Metformin in Human Plasma Using an Ion-Pair LC Method

Automation of pH Optimization Experiments During LC Development

The effect of pH on the high pH anion-exchange chromatography elution of monosaccharides

Amino-Functionalized Monolithic Poly(glycidyl methacrylate-co-divinylbenzene) Ion-Exchange Stationary Phases for the Separation of Oligonucleotides

The Behaviour of Reduced, Alkylated and Native Proteins in a pH-Gradient LC System

Similar Documents

Refolding human lysozyme produced as an inclusion body by urea concentration and pH gradient ion exchange chromatography

Investigation of the ion-exchange behaviour of titania: Application as a packing material for ion chromatography

Retention behaviour of an homologous series of oligodeoxythymidilic acids using reversed-phase ion-pair chromatography

LC–MS–MS Determination of Olmesartan in Human Plasma

Gas chromatographic behaviour of urea herbicides

Simple and Sensitive Determination of Metformin in Human Plasma Using an Ion-Pair LC Method

Automation of pH Optimization Experiments During LC Development

The effect of pH on the high pH anion-exchange chromatography elution of monosaccharides

Amino-Functionalized Monolithic Poly(glycidyl methacrylate-co-divinylbenzene) Ion-Exchange Stationary Phases for the Separation of Oligonucleotides

The Behaviour of Reduced, Alkylated and Native Proteins in a pH-Gradient LC System